Na⁺/K⁺-ATPase-Targeted Cytotoxicity of (+)-Digoxin and Several Semisynthetic Derivatives

Abstract

(+)-Digoxin (<b>1</b>) is a well-known cardiac glycoside long used to treat congestive heart failure and found more recently to show anticancer activity. Several known cardenolides (<b>2</b>-<b>5</b>) and two new analogues, (+)-8(9)-β-anhydrodigoxigenin (<b>6</b>) and (+)-17-<i>epi</i>-20,22-dihydro-21α-hydroxydigoxin (<b>7</b>), were synthesized from <b>1</b> and evaluated for their cytotoxicity toward a small panel of human cancer cell lines. A preliminary structure-activity relationship investigation conducted indicated that the C-12 and C-14 hydroxy groups and the C-17 unsaturated lactone unit are important for <b>1</b> to mediate its cytotoxicity toward human cancer cells, but the C-3 glycosyl residue seems to be less critical for such an effect. Molecular docking profiles showed that the cytotoxic <b>1</b> and the noncytotoxic derivative <b>7</b> bind differentially to Na⁺/K⁺-ATPase. The HO-12β, HO-14β, and HO-3'aα hydroxy groups of (+)-digoxin (<b>1</b>) may form hydrogen bonds with the side-chains of Asp121 and Asn122, Thr797, and Arg880 of Na⁺/K⁺-ATPase, respectively, but the altered lactone unit of <b>7</b> results in a rotation of its steroid core, which depotentiates the binding between this compound and Na⁺/K⁺-ATPase. Thus, <b>1</b> was found to inhibit Na⁺/K⁺-ATPase, but <b>7</b> did not. In addition, the cytotoxic <b>1</b> did not affect glucose uptake in human cancer cells, indicating that this cardiac glycoside mediates its cytotoxicity by targeting Na⁺/K⁺-ATPase but not by interacting with glucose transporters.